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. 2013 Apr 30;21(6):1232–1241. doi: 10.1038/mt.2013.59

Figure 4.

Figure 4

Immune responses to B2M-targeted cells. (a) MLR results showing 3[H]-thymidine incorporation by PBMC responder cells mixed with irradiated, undifferentiated B2M+/+ or B2MloxP/loxP ESCs (left panel) or with day 15 EB-derived cells (right panel) at a ratio of 1:1. Two independent PBMC responder cell preparations were used when analyzing EB cells. Controls included unstimulated PBMCs, and PBMCs stimulated by allogeneic PBMCs. (b) Flow cytometry analysis of day 15 B2M+/+ or B2MloxP/loxP EB cells showing the surface expression of CD34, HLA class I (HLA-A, -B, and -C), and HLA class II (HLA-DR), with isotype controls. All preparations were labeled with 7-amino-actinomycin D (7AAD) to improve gating and remove dead cells. (c) Intracellular cytokine staining of HLA-A*0201/NLV-CMVpp65-specific T cells stimulated with an equal number of B2M+/+ or B2MloxP/loxP EB cells, HLA class I–deficient K562 cells, or HLA-A*0201–expressing K562-A2 cells, with or without prior pulsing with NLVPMVATV (NLV) peptide from the CMV pp65 protein. HLA, human leukocyte antigens.